Monoclonal antibodies (MAbs) have revolutionized the conduct of science since their first description in 1975. The use of these specific reagents also has made possible improved clinical diagnostics in the medical arena, and a few antibodies have found their way to clinical use as prophylactic or therapeutic agents. Nevertheless, the potential of MAbs for therapy remains largely unfulfilled. The principal reason for the lack of a large number of MAb therapeutics is simply the difficulty in generating human monoclonal antibodies of high affinity. The objective of this proposal is to develop novel methods for the rapid and efficient generation of human B cell hybridomas secreting antibodies of medical importance. The work will develop and employ new approaches to rare antigen-specific B cell sorting from memory cell populations, and will result in the design and manufacture of new cell-cell electrofusion hardware and protocols.Three specific aims are proposed: 1) To develop high-throughput methods for antigen-specific human B cell physical sorting and expansion; 2) To develop novel electrofusion devices and protocols that yield a high frequency of viable human hybridomas: 3) To develop high affinity neutralizing antibodies to respiratory syncytial virus, a medically important viral pathogen amenable to MAb prophylaxis, as a demonstration of the feasibility of making important antiviral antibodies using the novel techniques and equipment developed.